Human Corticotropin Releasing Hormone ELISA Kit

Distributed by Antibodies.com, this competitive Enzyme-Linked Immunosorbent Assay (cELISA) is intended for the in vitro quantitative determination of human CRF (Corticotropin Releasing Hormone) in serum, plasma or other biological fluids. The 96-well microtiter plate is pre-coated with CRF antigen; during incubation, CRF present in samples or standards competes with a fixed amount of immobilized CRF for binding sites on the Biotinylated Anti-CRF Antibody. Following washing, an HRP-Avidin conjugate is added and TMB substrate solution is used to visualise the enzymatic reaction as a blue colour that changes to yellow after the addition of an acidic stop solution. Yellow intensity is inversely proportional to CRF concentration, which is calculated by reading O.D. absorbance at 450 nm against a standard curve. The platform is a pre-coated microplate (12 x 8 well strips) with a total assay time of 2h 30m, using a Colorimetric Microplate Reader.

Reported performance figures include an assay range of 15.6-1000 pg/ml and a sensitivity of <4.24 pg/ml. Spiking recovery data with n=5 samples showed ranges, with averages in parentheses, of 80%-102% (91%) for Serum, 81%-100% (90%) for EDTA Plasma and 80%-89% (84%) for Heparin Plasma. Dilution linearity at 1:2, 1:4, 1:8 and 1:16 yielded recoveries of 87-91%, 87-107%, 74-101% and 92-97% for Serum (n=5); 90-105%, 84-101%, 90-101% and 79-108% for EDTA Plasma (n=5); and 84-95%, 92-105%, 82-105% and 89-91% for Heparin Plasma (n=5).

Kit components are stored at +4°C or -20°C as directed, and the kit must not be used past its expiration date. Components supplied include a Pre-Coated 96 Well Microplate, Lyophilized Standard, Detection Solution A, Detection Solution B, Wash Buffer (30X), Sample Dilution Buffer, TMB Substrate, Stop Solution and Plate Sealers (Adhesive Strips). The product falls within the Antibodies.com Assays and Kits category.